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A liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assay to profile 20 plasma steroids in endocrine disorders

  • Zhenxin Wang , Hao Wang ORCID logo , Yingfei Peng , Fangjun Chen ORCID logo , Lin Zhao , Xiaomu Li ORCID logo , Jiaqian Qin , Qianqian Li , Beili Wang , Baishen Pan and Wei Guo EMAIL logo
Published/Copyright: February 21, 2020
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 58 Issue 9

Abstract

Background

Liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assays are employed in more and more clinical laboratories to quantify steroids. The steroid quantification by LC-MS/MS shows great value in screening or diagnosing endocrine disorders; however, the number of functional steroids included in the LC-MS/MS methods is still limited.

Methods

Here, we describe the performance and validation of a 20-steroid plasma panel by LC-MS/MS. The panel included progestogens (including mineralocorticoids and glucocorticoids), androgens and estrogens biosynthesized in steroid metabolic pathways. The LC-MS/MS method was validated according to guidance documents, and subsequently employed to profile steroid changes in endocrine disorders.

Results

Using LC-MS/MS, 20 steroids were separated and quantified in 8 min. Coefficients of variation (CVs) of the 20 analytes at the lower limit of quantification (LLoQ) were all less than 15% (ranging from 1.84% to 14.96%). The linearity of the assay was demonstrated by all the R2 values greater than 0.995. Individual plasma steroids changed significantly in patients with subclinical Cushing’s syndrome (SCS) and polycystic ovary syndrome (PCOS) – 17-hydroxypregnenolone (17-OH-PR), testosterone (T) and dihydrotestosterone (DHT) were significantly decreased in SCS patients, while in PCOS patients, pregnenolone, corticosterone (CORT), androstenedione (A4) and T were significantly increased and DHT was decreased.

Conclusions

The LC-MS/MS method we developed for the quantification of 20 plasma steroids is clinical practicable. The steroid profiling data using this assay indicate its screening value for endocrine disorders. To further explore the value of the assay, more investigations are however needed.

Keywords: endocrine disorders; liquid chromatography-tandem mass spectrometry (LC-MS/MS); panel; pathway; steroids
Corresponding author: Wei Guo, PhD, Department of Laboratory Medicine, Zhongshan Hospital, Fudan University, 136 Yi Xue Yuan Road, Shanghai 200032, P.R. China
aZhenxin Wang, Hao Wang, Yingfei Peng and Fangjun Chen contributed equally to this work.

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission. W.G. made the concept. W.G., B.W. and Z.W. contributed to the study design. Y.P. and F.C. contributed to method development and method validation. L.Z. and X.L. contributed to classification and collection of clinical samples. J.Q. contributed to data acquisition. Z.W. and H.W. contributed to analysis and interpretation of the data. Z.W. and H.W. contributed to the drafting of the manuscript. H.W., B.P. and W.G. contributed to the critical revision of the manuscript. Q.L. contributed to technology support.

  2. Research funding: Wei Guo was supported by Joint Project of Fudan University & Suzhou Institute of Biomedical Engineering and Technology, Chinese Academy of Sciences, the National Natural Science Foundation of China (81772263, 81972000 and 81572064), Shanghai Municipal Key Clinical Specialty and Key Developing Disciplines of Shanghai Municipal Commission of Health and Family Planning (2015ZB0201). Hao Wang was supported by Shanghai Post-doctoral Excellence Program (2018166) and the project funded by China Postdoctoral Science Foundation (2019M651370, Funder Id: http://dx.doi.org/10.13039/501100002858). Beili Wang was supported by the Sponsorship for the Junior Clinical Medical Technologist in Shanghai (201802), the National Natural Science Foundation of China (81902139) and the Projects from Shanghai Science and Technology Commission (16411952100).

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2019-0869).

Received: 2019-08-16
Accepted: 2020-01-02
Published Online: 2020-02-21
Published in Print: 2020-08-27

©2020 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2019-0869
Keywords for this article
endocrine disorders; liquid chromatography-tandem mass spectrometry (LC-MS/MS); panel; pathway; steroids

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  1. Frontmatter
  2. Editorial
  3. Measurement uncertainty: light in the shadows
  4. Review
  5. Childhood obesity: an overview of laboratory medicine, exercise and microbiome
  6. Mini review
  7. The utility of measurement uncertainty in medical laboratories
  8. Opinion Paper
  9. Do genetic polymorphisms in angiotensin converting enzyme 2 (ACE2) gene play a role in coronavirus disease 2019 (COVID-19)?
  10. Opinion Papers
  11. The current status and future prospects of precision medicine
  12. Precision medicine in medical oncology: hope, disappointment and reality
  13. IFCC Papers
  14. Laboratory practices to mitigate biohazard risks during the COVID-19 outbreak: an IFCC global survey
  15. Operational considerations and challenges of biochemistry laboratories during the COVID-19 outbreak: an IFCC global survey
  16. Genetics and Molecular Diagnostics
  17. Diverse fragment lengths dismiss size selection for serum cell-free DNA: a comparative study of serum and plasma samples
  18. General Clinical Chemistry and Laboratory Medicine
  19. Quantification of plasma remdesivir and its metabolite GS-441524 using liquid chromatography coupled to tandem mass spectrometry. Application to a Covid-19 treated patient
  20. Isotope dilution-LC-MS/MS method for quantification of the urinary cotinine-to-creatinine ratio
  21. A liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assay to profile 20 plasma steroids in endocrine disorders
  22. Assessment of antinuclear antibodies by indirect immunofluorescence assay: report from a survey by the American Association of Medical Laboratory Immunologists
  23. Evaluation of a novel extended automated particle-based multi-analyte assay for the detection of autoantibodies in the diagnosis of primary biliary cholangitis
  24. Reference Values and Biological Variations
  25. Continuous, complete and comparable NT-proBNP reference ranges in healthy children
  26. Reference-mean-centered statistical quality control
  27. Hematology and Coagulation
  28. Top-down and bottom-up approaches for the estimation of measurement uncertainty in coagulation assays
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